Date of Award

9-2004

Degree Type

Thesis

Degree Name

MS Biology

Department

Biology

Advisor

Jane L. Ko

Committee Member

Carolyn Bentivenga

Committee Member

Allan D. Blake

Committee Member

Sulie Chang

Keywords

KH domains, Poly C binding protein, Mu-opiod receptor gene

Abstract

The mu-opioid receptor (MOR) has been shown to mediate morphine induced analgesia, addiction, and tolerance. The expression of the major product of the MOR (MOR-1) gene has been found to be driven by two promoters, a distal promoter and a proximal promoter. The proximal promoter region has been shown in previous studies to be the major promoter for the regulation of MOR gene transcription in the adult mouse brain and embryonic development. Recent studies have shown that single-stranded DNA binding proteins (such as Poly C Binding Protein) are involved in transcriptional regulation of mouse MOR gene. Poly C binding protein (PCBPs) is categorized in the family of hnRNP's, containing multiple K-Homology (KH) domains, which is involved in RNA stabilization, translational activation, and gene silencing. In this study, we attempted to determine the trans-activation domains of PCBP and examine the regulatory effects of each trans-activation domain of PCBP in MOR transcription. Mouse neuronal cells (N2A) were transiently transfected with various truncation constructs of PCBP to determine which domains were important in trans­ activation. Our preliminary data suggests that at least two KH domains were required for PCBP to display its trans-activation activity.

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